https://padlet.com/nthorogood/6zptdka0br5s en-us 2019-12-09 13:29:31 UTC 2025-04-05 14:36:36 UTC hello@padlet.com nthorogood https://padlet.com/nthorogood/6zptdka0br5s/wish/421572230 2019-12-09 13:33:11 UTC https://padlet.com/nthorogood/6zptdka0br5s/wish/421572230 nthorogood https://padlet.com/nthorogood/6zptdka0br5s/wish/421573193 2019-12-09 13:35:15 UTC https://padlet.com/nthorogood/6zptdka0br5s/wish/421573193 nthorogood https://padlet.com/nthorogood/6zptdka0br5s/wish/421577278 The overall purpose of the Protein Factories lab is to transform and purify Green Fluorescent Protein (GFP) in bacteria.]]> 2019-12-09 13:42:53 UTC https://padlet.com/nthorogood/6zptdka0br5s/wish/421577278 nthorogood https://padlet.com/nthorogood/6zptdka0br5s/wish/422200289 Purpose: The purpose of bacterial transformation is to introduce a foreign plasmid into bacteria.  The bacteria can then amplify the plasmid and make large quantities of it.]]> 2019-12-10 15:12:40 UTC https://padlet.com/nthorogood/6zptdka0br5s/wish/422200289 nthorogood https://padlet.com/nthorogood/6zptdka0br5s/wish/422200599 Purpose: The purpose of protein purification is to separate the protein of interest from other materials, such as cell particles or other proteins.  This process is vital for the characterization of the function, structure, and interactions for the desired protein (in this lab, the GFP protein).]]> 2019-12-10 15:13:05 UTC https://padlet.com/nthorogood/6zptdka0br5s/wish/422200599 nthorogood https://padlet.com/nthorogood/6zptdka0br5s/wish/422200892 Purpose:  The reason why protein gel electrophoresis is being used in this lab is so that the GFP samples (both the native and denatured) can be analyzed.]]> 2019-12-10 15:13:30 UTC https://padlet.com/nthorogood/6zptdka0br5s/wish/422200892 nthorogood https://padlet.com/nthorogood/6zptdka0br5s/wish/422213577 The first step in the transformation process is to add calcium chloride (CaCl2) to the bacteria cell. This will allow for the bacteria to incorporate it´s plasmid DNA and allow for an easy transformation process.]]> 2019-12-10 15:29:31 UTC https://padlet.com/nthorogood/6zptdka0br5s/wish/422213577 nthorogood https://padlet.com/nthorogood/6zptdka0br5s/wish/422221420 The next part of the transformation process is to twist an inoculating loop to free all the cells and make sure each cell is transformed with the GFP Protein. ]]> 2019-12-10 15:39:17 UTC https://padlet.com/nthorogood/6zptdka0br5s/wish/422221420 nthorogood https://padlet.com/nthorogood/6zptdka0br5s/wish/422233904 Transfer 250 micro liters of the cell suspension to the ¨+DNA¨ tube and place both tubes on ice.]]> 2019-12-10 15:54:45 UTC https://padlet.com/nthorogood/6zptdka0br5s/wish/422233904 nthorogood https://padlet.com/nthorogood/6zptdka0br5s/wish/422309904 Incubate the tubes on ice for 10 minutes so that bacteria can interact with the calcium ion.]]> 2019-12-10 17:44:28 UTC https://padlet.com/nthorogood/6zptdka0br5s/wish/422309904 nthorogood https://padlet.com/nthorogood/6zptdka0br5s/wish/422313933 Once the bacteria has incubated on the ice for 10 minutes, place the transformation tubes in a 42 deg C water bath for 45 seconds. The purpose of putting the bacteria colonies in the water bath is to allow the plasmid DNA to interact with the bacteria.]]> 2019-12-10 17:50:11 UTC https://padlet.com/nthorogood/6zptdka0br5s/wish/422313933 nthorogood https://padlet.com/nthorogood/6zptdka0br5s/wish/422924775 The main structures of a pFluorogreen Plasmid include the following genes.]]> 2019-12-11 19:08:00 UTC https://padlet.com/nthorogood/6zptdka0br5s/wish/422924775 nthorogood https://padlet.com/nthorogood/6zptdka0br5s/wish/422927933 Bla (beta-lactamase enzyme) allows for bacteria to become resistant to the beta-lactam class of antibiotics (penicillin class of antibiotics).]]> 2019-12-11 19:11:46 UTC https://padlet.com/nthorogood/6zptdka0br5s/wish/422927933 nthorogood https://padlet.com/nthorogood/6zptdka0br5s/wish/422930317 araC is another important part of the pFluorogreen Plasmid, which is in charge of the expression of the GFP Protein.]]> 2019-12-11 19:15:03 UTC https://padlet.com/nthorogood/6zptdka0br5s/wish/422930317 nthorogood https://padlet.com/nthorogood/6zptdka0br5s/wish/422938690 The main functions of the GFP Protein are to tag proteins and to examine cellular processes within living cells.]]> 2019-12-11 19:26:30 UTC https://padlet.com/nthorogood/6zptdka0br5s/wish/422938690 nthorogood https://padlet.com/nthorogood/6zptdka0br5s/wish/423811353 After the bacteria has been in the water bath for 45 seconds, place the bacteria back on the ice for 2 minutes so that the bacteria can continue to incubate and interact with the calcium chloride.]]> 2019-12-13 14:40:04 UTC https://padlet.com/nthorogood/6zptdka0br5s/wish/423811353 nthorogood https://padlet.com/nthorogood/6zptdka0br5s/wish/423817138 Once the bacterial cells have incubated on the ice for 2 minutes, 250 micro liters of recovery broth must be transferred in order to promote improved cell viability and greater cloning efficiency. ]]> 2019-12-13 14:48:08 UTC https://padlet.com/nthorogood/6zptdka0br5s/wish/423817138 nthorogood https://padlet.com/nthorogood/6zptdka0br5s/wish/423823571 Once recovery broth has been added to the bacterial tubes, incubate the cells in a 37 deg C water bath for 10 minutes to continue to allow the plasmid DNA to interact with the bacteria.]]> 2019-12-13 14:57:27 UTC https://padlet.com/nthorogood/6zptdka0br5s/wish/423823571 nthorogood https://padlet.com/nthorogood/6zptdka0br5s/wish/424186496 2019-12-14 21:11:48 UTC https://padlet.com/nthorogood/6zptdka0br5s/wish/424186496 nthorogood https://padlet.com/nthorogood/6zptdka0br5s/wish/424186558 The techniques that are required to create a pFlurogreen plasmid are...]]> 2019-12-14 21:12:26 UTC https://padlet.com/nthorogood/6zptdka0br5s/wish/424186558 nthorogood https://padlet.com/nthorogood/6zptdka0br5s/wish/424186660 2019-12-14 21:13:43 UTC https://padlet.com/nthorogood/6zptdka0br5s/wish/424186660 nthorogood https://padlet.com/nthorogood/6zptdka0br5s/wish/424186683 2019-12-14 21:13:58 UTC https://padlet.com/nthorogood/6zptdka0br5s/wish/424186683 nthorogood https://padlet.com/nthorogood/6zptdka0br5s/wish/424186707 2019-12-14 21:14:15 UTC https://padlet.com/nthorogood/6zptdka0br5s/wish/424186707 nthorogood https://padlet.com/nthorogood/6zptdka0br5s/wish/424191667 2019-12-14 22:11:45 UTC https://padlet.com/nthorogood/6zptdka0br5s/wish/424191667 nthorogood https://padlet.com/nthorogood/6zptdka0br5s/wish/424192761 2019-12-14 22:24:31 UTC https://padlet.com/nthorogood/6zptdka0br5s/wish/424192761 nthorogood https://padlet.com/nthorogood/6zptdka0br5s/wish/424194096 Purpose: The purpose of isolating the GFP protein is so that the GFP can be separated from other materials, such as molecules or other proteins.]]> 2019-12-14 22:30:09 UTC https://padlet.com/nthorogood/6zptdka0br5s/wish/424194096 nthorogood https://padlet.com/nthorogood/6zptdka0br5s/wish/424195178 Using a sterile 1 ml pipet, transfer 250 micro liters of recovered cells from the tube labeled ¨-DNA¨ to the middle of the -DNA and
-DNA and -DNA/+Amp plates.]]> 2019-12-14 22:40:00 UTC https://padlet.com/nthorogood/6zptdka0br5s/wish/424195178 nthorogood https://padlet.com/nthorogood/6zptdka0br5s/wish/424196943 2019-12-14 22:54:37 UTC https://padlet.com/nthorogood/6zptdka0br5s/wish/424196943 nthorogood https://padlet.com/nthorogood/6zptdka0br5s/wish/424198756 2019-12-14 23:14:03 UTC https://padlet.com/nthorogood/6zptdka0br5s/wish/424198756 nthorogood https://padlet.com/nthorogood/6zptdka0br5s/wish/424199140 2019-12-14 23:18:24 UTC https://padlet.com/nthorogood/6zptdka0br5s/wish/424199140 nthorogood https://padlet.com/nthorogood/6zptdka0br5s/wish/424199663 2019-12-14 23:25:11 UTC https://padlet.com/nthorogood/6zptdka0br5s/wish/424199663 nthorogood https://padlet.com/nthorogood/6zptdka0br5s/wish/424199784 2019-12-14 23:26:53 UTC https://padlet.com/nthorogood/6zptdka0br5s/wish/424199784 nthorogood https://padlet.com/nthorogood/6zptdka0br5s/wish/424199999 2019-12-14 23:29:47 UTC https://padlet.com/nthorogood/6zptdka0br5s/wish/424199999 nthorogood https://padlet.com/nthorogood/6zptdka0br5s/wish/424200512 Using a new sterile 1 ml pipet, transfer 250 micro liters of recovered cells from the tube labeled ¨+DNA¨ to the middle of the +DNA/+Amp and +DNA/+Amp plates.]]> 2019-12-14 23:37:25 UTC https://padlet.com/nthorogood/6zptdka0br5s/wish/424200512 nthorogood https://padlet.com/nthorogood/6zptdka0br5s/wish/424200516 Spread the cells over the entire plate using an inoculating loop. Use one sterile loop to spread both -DNA samples. Change to a fresh loop before spreading the +DNA samples. The cells are being spread over the plates so that the cell suspensions will be absorbed by the agar solution and to promote the bacteria to grow.]]> 2019-12-14 23:37:30 UTC https://padlet.com/nthorogood/6zptdka0br5s/wish/424200516 nthorogood https://padlet.com/nthorogood/6zptdka0br5s/wish/424201626 GFP was originally discovered from the Aequorea Victoria species of jellyfish.]]> 2019-12-14 23:49:37 UTC https://padlet.com/nthorogood/6zptdka0br5s/wish/424201626 nthorogood https://padlet.com/nthorogood/6zptdka0br5s/wish/424209263 After performing these steps, the transformed bacteria should look like the bacteria on the right side of this image.]]> 2019-12-15 01:14:41 UTC https://padlet.com/nthorogood/6zptdka0br5s/wish/424209263 nthorogood https://padlet.com/nthorogood/6zptdka0br5s/wish/424212582 After performing gel electrophoresis on the GFP protein samples, the results should look like this picture.]]> 2019-12-15 01:45:32 UTC https://padlet.com/nthorogood/6zptdka0br5s/wish/424212582 nthorogood https://padlet.com/nthorogood/6zptdka0br5s/wish/424212961 Click to see image.]]> 2019-12-15 01:49:17 UTC https://padlet.com/nthorogood/6zptdka0br5s/wish/424212961 nthorogood https://padlet.com/nthorogood/6zptdka0br5s/wish/424213341 Click to see image.]]> 2019-12-15 01:52:31 UTC https://padlet.com/nthorogood/6zptdka0br5s/wish/424213341 nthorogood https://padlet.com/nthorogood/6zptdka0br5s/wish/424214105 With an inoculating loop, pick 4-5 isolated GFP expressing (glowing) colonies to isolate the colonies that have the GFP Protein.]]> 2019-12-15 01:59:34 UTC https://padlet.com/nthorogood/6zptdka0br5s/wish/424214105 nthorogood https://padlet.com/nthorogood/6zptdka0br5s/wish/424214867 Spread the cells evenly and thoroughly over the entire surface of the plate to allow for even distribution of the cells. Turn the plate 90 degrees and thoroughly spread again using the same loop.]]> 2019-12-15 02:07:13 UTC https://padlet.com/nthorogood/6zptdka0br5s/wish/424214867 nthorogood https://padlet.com/nthorogood/6zptdka0br5s/wish/424215331 Repeat the same process as steps 1 and 2 for the second plate.]]> 2019-12-15 02:11:25 UTC https://padlet.com/nthorogood/6zptdka0br5s/wish/424215331 nthorogood https://padlet.com/nthorogood/6zptdka0br5s/wish/424215522 Incubate the plates in a 37 degree incubator overnight.]]> 2019-12-15 02:12:56 UTC https://padlet.com/nthorogood/6zptdka0br5s/wish/424215522 nthorogood https://padlet.com/nthorogood/6zptdka0br5s/wish/424215669 Select a GFP plate showing the highest GFP expression (maximum glow). Using a sterile loop, scrape the cell growth off of the GFP plate to further isolate the cells.]]> 2019-12-15 02:14:21 UTC https://padlet.com/nthorogood/6zptdka0br5s/wish/424215669 nthorogood https://padlet.com/nthorogood/6zptdka0br5s/wish/424215973 Twirl the loop containing the colonies in the tube containing lysis buffer. The purpose of twirling is to dislodge the cells into the buffer solution.]]> 2019-12-15 02:17:21 UTC https://padlet.com/nthorogood/6zptdka0br5s/wish/424215973 nthorogood https://padlet.com/nthorogood/6zptdka0br5s/wish/424216534 Vortex the bacterial tube at maximum speed. The purpose of vortexing is to make sure the cells are fully resuspended.]]> 2019-12-15 02:22:59 UTC https://padlet.com/nthorogood/6zptdka0br5s/wish/424216534 nthorogood https://padlet.com/nthorogood/6zptdka0br5s/wish/424217150 Incubate tube for 10 minutes in a 55 degree Celsius bath.]]> 2019-12-15 02:28:11 UTC https://padlet.com/nthorogood/6zptdka0br5s/wish/424217150 nthorogood https://padlet.com/nthorogood/6zptdka0br5s/wish/424217450 Place the microcentrifuge tube containing the GFP cells in a -20 degree Celsius freezer for 15 minutes.]]> 2019-12-15 02:30:46 UTC https://padlet.com/nthorogood/6zptdka0br5s/wish/424217450 nthorogood https://padlet.com/nthorogood/6zptdka0br5s/wish/424217937 After the cell suspension is completely frozen, remove the centrifuge tube from the freezer and put it in a 55 degree Celsius water bath to thaw the cells.]]> 2019-12-15 02:34:09 UTC https://padlet.com/nthorogood/6zptdka0br5s/wish/424217937 nthorogood https://padlet.com/nthorogood/6zptdka0br5s/wish/424218218 Vortex the samples vigorously for 30 seconds.]]> 2019-12-15 02:36:26 UTC https://padlet.com/nthorogood/6zptdka0br5s/wish/424218218 nthorogood https://padlet.com/nthorogood/6zptdka0br5s/wish/424218395 Repeat steps 9-12 two more times. The purpose of the freezing, melting, and vortexing pattern is to help lyse the bacterial cells.]]> 2019-12-15 02:37:51 UTC https://padlet.com/nthorogood/6zptdka0br5s/wish/424218395 nthorogood https://padlet.com/nthorogood/6zptdka0br5s/wish/424218827 Centrifuge the tube in a microcentrifuge for 5 minutes to further lyse the cells.]]> 2019-12-15 02:42:05 UTC https://padlet.com/nthorogood/6zptdka0br5s/wish/424218827 nthorogood https://padlet.com/nthorogood/6zptdka0br5s/wish/424228931 Click to see image.]]> 2019-12-15 04:21:49 UTC https://padlet.com/nthorogood/6zptdka0br5s/wish/424228931 nthorogood https://padlet.com/nthorogood/6zptdka0br5s/wish/424229037 Click to see image.]]> 2019-12-15 04:22:51 UTC https://padlet.com/nthorogood/6zptdka0br5s/wish/424229037 nthorogood https://padlet.com/nthorogood/6zptdka0br5s/wish/424229934 Vertically mount the column on a ring stand. Make sure it is straight and that the white cap is firmly attached to the bottom of the column.]]> 2019-12-15 04:34:49 UTC https://padlet.com/nthorogood/6zptdka0br5s/wish/424229934 nthorogood https://padlet.com/nthorogood/6zptdka0br5s/wish/424232013 Add 1 ml of 1X elution buffer to the column to prevent the column from drying.]]> 2019-12-15 04:59:31 UTC https://padlet.com/nthorogood/6zptdka0br5s/wish/424232013 nthorogood https://padlet.com/nthorogood/6zptdka0br5s/wish/424233702 Pipet 4 ml of the mixed slurry into the column by letting it stream down the inside walls of the column. The purpose of adding slurry is to have more efficiency in separating the green fluorescent protein.]]> 2019-12-15 05:18:22 UTC https://padlet.com/nthorogood/6zptdka0br5s/wish/424233702 nthorogood https://padlet.com/nthorogood/6zptdka0br5s/wish/424363462 Place an empty beaker under the column in order to collect the 1X elution buffer. ]]> 2019-12-16 01:39:34 UTC https://padlet.com/nthorogood/6zptdka0br5s/wish/424363462 nthorogood https://padlet.com/nthorogood/6zptdka0br5s/wish/424363935 Once the beaker is placed underneath the column, remove the cap from the bottom of the column so that the matrix can be packed into the column]]> 2019-12-16 01:42:56 UTC https://padlet.com/nthorogood/6zptdka0br5s/wish/424363935 nthorogood https://padlet.com/nthorogood/6zptdka0br5s/wish/424371241 Wash the packed column with 5 ml of 1X elution buffer. Always keep a thin layer of elution buffer on top of the packed matrix to prevent the solution from drying.]]> 2019-12-16 02:33:37 UTC https://padlet.com/nthorogood/6zptdka0br5s/wish/424371241 nthorogood https://padlet.com/nthorogood/6zptdka0br5s/wish/424372059 Slowly load the column with 0.2 ml of the GFP extract.]]> 2019-12-16 02:38:50 UTC https://padlet.com/nthorogood/6zptdka0br5s/wish/424372059 nthorogood https://padlet.com/nthorogood/6zptdka0br5s/wish/424372649 Begin to elute the column with 1X elution buffer. Add the buffer slowly. The purpose of eluting the column with 1X elution buffer is to avoid diluting the protein sample. ]]> 2019-12-16 02:43:13 UTC https://padlet.com/nthorogood/6zptdka0br5s/wish/424372649 nthorogood https://padlet.com/nthorogood/6zptdka0br5s/wish/424372739 Remove the cap so that the extract can completely enter the column, collecting the ¨flow-through¨ waste into the beaker.]]> 2019-12-16 02:43:50 UTC https://padlet.com/nthorogood/6zptdka0br5s/wish/424372739 nthorogood https://padlet.com/nthorogood/6zptdka0br5s/wish/424373570 Once these steps are accomplished, the bacterial cells should be lysed like the cells in the microscopic diagram.]]> 2019-12-16 02:48:44 UTC https://padlet.com/nthorogood/6zptdka0br5s/wish/424373570 nthorogood https://padlet.com/nthorogood/6zptdka0br5s/wish/424376808 GFP is related to diabetes and insulin because the protein acts as an imaging source to determine whether or not enough insulin is being secreted by the pancreas and into the cells.]]> 2019-12-16 03:09:08 UTC https://padlet.com/nthorogood/6zptdka0br5s/wish/424376808 nthorogood https://padlet.com/nthorogood/6zptdka0br5s/wish/424377078 By using GFP as an imaging source, if the GFP test shows that the pancreas is not making enough insulin for the cells, doctors can help better diagnose and treat diabetic patients.]]> 2019-12-16 03:11:22 UTC https://padlet.com/nthorogood/6zptdka0br5s/wish/424377078 nthorogood https://padlet.com/nthorogood/6zptdka0br5s/wish/424384900 When the GFP Protein band almost reaches the bottom of the column, start collecting the fractions in the microtiter plate. In order to prevent any loss of GFP protein, begin collecting fractions before the GFP reaches the bottom of the column.]]> 2019-12-16 04:05:25 UTC https://padlet.com/nthorogood/6zptdka0br5s/wish/424384900 nthorogood https://padlet.com/nthorogood/6zptdka0br5s/wish/424385845 Transfer 30 micro liters of the brightest elution to a screw-top microcentrifuge tube. This tube will be labeled ¨GFP Native.¨ ]]> 2019-12-16 04:14:19 UTC https://padlet.com/nthorogood/6zptdka0br5s/wish/424385845 nthorogood https://padlet.com/nthorogood/6zptdka0br5s/wish/424387019 Transfer an additional 30 micro liters of the same elution to a second screw-top microcentrifuge tube. This tube will be labeled ¨GFP Denatured.¨]]> 2019-12-16 04:22:29 UTC https://padlet.com/nthorogood/6zptdka0br5s/wish/424387019 nthorogood https://padlet.com/nthorogood/6zptdka0br5s/wish/424388018 The purified GFP Protein will look like the image below.]]> 2019-12-16 04:29:13 UTC https://padlet.com/nthorogood/6zptdka0br5s/wish/424388018 nthorogood https://padlet.com/nthorogood/6zptdka0br5s/wish/424390978 Click to see image.]]> 2019-12-16 04:53:36 UTC https://padlet.com/nthorogood/6zptdka0br5s/wish/424390978 nthorogood https://padlet.com/nthorogood/6zptdka0br5s/wish/424391062 Click to see image.]]> 2019-12-16 04:54:27 UTC https://padlet.com/nthorogood/6zptdka0br5s/wish/424391062 nthorogood https://padlet.com/nthorogood/6zptdka0br5s/wish/424391079 Click to see image.]]> 2019-12-16 04:54:38 UTC https://padlet.com/nthorogood/6zptdka0br5s/wish/424391079 nthorogood https://padlet.com/nthorogood/6zptdka0br5s/wish/424391093 Click to see image.]]> 2019-12-16 04:54:47 UTC https://padlet.com/nthorogood/6zptdka0br5s/wish/424391093 nthorogood https://padlet.com/nthorogood/6zptdka0br5s/wish/424712780 The purpose of denaturing proteins is to prevent the loss of the protein´s function (in this case, the GFP protein). To denature the protein sample, add 10 micro liters of protein denaturing solution to the tube labeled ¨GFP denatured¨ and mix well. The denaturing solution contains sodium dodecyl sulfate (SDS) and 2-mercaptoethanol.]]> 2019-12-16 19:31:18 UTC https://padlet.com/nthorogood/6zptdka0br5s/wish/424712780 nthorogood https://padlet.com/nthorogood/6zptdka0br5s/wish/424752914 Push the bottom of the GFP sample tube through the foil and immerse in the boiling water. The protein in the protein denaturing solution, along with the boiling water, will help denature the GFP protein.]]> 2019-12-16 20:54:48 UTC https://padlet.com/nthorogood/6zptdka0br5s/wish/424752914 nthorogood https://padlet.com/nthorogood/6zptdka0br5s/wish/424755623 Boil the GFP sample for 5 minutes. Afterwards, remove the sample tube from the beaker and allow it to cool for a few minutes at room temperature.]]> 2019-12-16 21:01:30 UTC https://padlet.com/nthorogood/6zptdka0br5s/wish/424755623 nthorogood https://padlet.com/nthorogood/6zptdka0br5s/wish/424756711 Gels may feature a sticker or tape at the bottom of the front plate. Remove the tape if present so that the bottom of the gel will be exposed.]]> 2019-12-16 21:03:58 UTC https://padlet.com/nthorogood/6zptdka0br5s/wish/424756711 nthorogood https://padlet.com/nthorogood/6zptdka0br5s/wish/424759102 Carefully remove the comb by gently pulling upwards. Pull the comb straight up to prevent damage to the wells.]]> 2019-12-16 21:10:15 UTC https://padlet.com/nthorogood/6zptdka0br5s/wish/424759102 nthorogood https://padlet.com/nthorogood/6zptdka0br5s/wish/424759975 Insert the gel into the electrophoresis chamber. The gel will promote the separation of the GFP protein´s molecules.]]> 2019-12-16 21:12:41 UTC https://padlet.com/nthorogood/6zptdka0br5s/wish/424759975 nthorogood https://padlet.com/nthorogood/6zptdka0br5s/wish/424761862 Add diluted electrophoresis buffer to the chamber.  The purpose of adding the buffer is transmit charge throughout the gel.]]> 2019-12-16 21:17:57 UTC https://padlet.com/nthorogood/6zptdka0br5s/wish/424761862 nthorogood https://padlet.com/nthorogood/6zptdka0br5s/wish/424763331 Rinse each well by squirting electrophoresis buffer into the wells using a transfer pipet. Once this step is done, the gel will now by ready for GFP protein sample loading because the buffer will now allow the GFP´s molecules to be separated by charge.]]> 2019-12-16 21:22:08 UTC https://padlet.com/nthorogood/6zptdka0br5s/wish/424763331 nthorogood https://padlet.com/nthorogood/6zptdka0br5s/wish/424765078 Perform gel electrophoresis on the GFP protein so that the GFP samples can be analyzed.]]> 2019-12-16 21:26:37 UTC https://padlet.com/nthorogood/6zptdka0br5s/wish/424765078 nthorogood https://padlet.com/nthorogood/6zptdka0br5s/wish/424768512 After gel electrophoresis has been performed, lay the cassette down and remove the front plate.]]> 2019-12-16 21:36:54 UTC https://padlet.com/nthorogood/6zptdka0br5s/wish/424768512 nthorogood https://padlet.com/nthorogood/6zptdka0br5s/wish/424769252 Add a sufficient volume (approximately 100 ml) of the staining/destaining solution into the tray to cover the gel and back plate.]]> 2019-12-16 21:39:24 UTC https://padlet.com/nthorogood/6zptdka0br5s/wish/424769252 nthorogood https://padlet.com/nthorogood/6zptdka0br5s/wish/424770083 Carefully remove the back plate from the tray.]]> 2019-12-16 21:42:27 UTC https://padlet.com/nthorogood/6zptdka0br5s/wish/424770083 nthorogood https://padlet.com/nthorogood/6zptdka0br5s/wish/424770536 Gently float a sheet of Protein InstaStain with the stain side (blue side) down in the staining/destaining solution. Cover the gel with plastic wrap to prevent evaporation.]]> 2019-12-16 21:44:12 UTC https://padlet.com/nthorogood/6zptdka0br5s/wish/424770536 nthorogood https://padlet.com/nthorogood/6zptdka0br5s/wish/424771400 Allow the Protein InstaStain paper to stain the gel for about an hour at room temperature.]]> 2019-12-16 21:47:25 UTC https://padlet.com/nthorogood/6zptdka0br5s/wish/424771400 nthorogood https://padlet.com/nthorogood/6zptdka0br5s/wish/424771875 Agitate on a rocking platform for 2-3 hours.]]> 2019-12-16 21:49:10 UTC https://padlet.com/nthorogood/6zptdka0br5s/wish/424771875 nthorogood https://padlet.com/nthorogood/6zptdka0br5s/wish/424792954 GFP is also related to insulin because insulin can be produced from the transcription of the bacterial cell´s plasmid.]]> 2019-12-16 23:19:16 UTC https://padlet.com/nthorogood/6zptdka0br5s/wish/424792954 nthorogood https://padlet.com/nthorogood/6zptdka0br5s/wish/424794601 This GFP from the jellyfish is able to be produced as a result of DNA Recombinant Technology. ]]> 2019-12-16 23:28:11 UTC https://padlet.com/nthorogood/6zptdka0br5s/wish/424794601 nthorogood https://padlet.com/nthorogood/6zptdka0br5s/wish/424796345 The GFP is made from a gene that is specifically designed to create GFP.]]> 2019-12-16 23:38:37 UTC https://padlet.com/nthorogood/6zptdka0br5s/wish/424796345 nthorogood https://padlet.com/nthorogood/6zptdka0br5s/wish/424797368 Once the GFP gene is produced, scientists can attach tags to selected proteins to monitor cellular activity in organisms.]]> 2019-12-16 23:44:28 UTC https://padlet.com/nthorogood/6zptdka0br5s/wish/424797368